Riddell Dominique O, Hildyard John C W, Harron Rachel C M, Wells Dominic J, Piercy Richard J
Comparative Neuromuscular Diseases Laboratory, Department of Clinical Science and Services, Royal Veterinary College, London, NW10TU, UK.
Department of Comparative Biomedical Sciences, Royal Veterinary College, London, NW10TU, UK.
Wellcome Open Res. 2022 Aug 17;6:354. doi: 10.12688/wellcomeopenres.17398.2. eCollection 2021.
Duchenne muscular dystrophy (DMD) is a fatal muscle wasting disease caused by mutations in the dystrophin gene. Due to their phenotypic similarity to human patients, large animal models are invaluable tools for pre-clinical trials. The DE50-MD dog is a relatively new model of DMD, and carries a therapeutically-tractable mutation lying within the hotspot for human patients, making it especially valuable. Prior to conducting therapeutic trials using this novel animal model, it is essential to establish a panel of viable biomarkers. We evaluated a panel of blood-borne biomarkers of musculoskeletal disease in the DE50-MD dog. Venous blood samples were obtained monthly throughout an 18-month study period in DE50-MD (N=18) and wild-type (WT) control (N=14) dogs. A panel of potential plasma/serum biomarkers of DMD was measured and their theoretical utility in future clinical trials determined using sample size calculations. Compared to WT dogs, DE50-MD dogs had substantially higher circulating creatine kinase (CK) activities, myomesin-3 (MYOM3), and the dystromiRs miR-1, miR-133a and miR-206, but significantly lower serum myostatin concentrations. An age-associated pattern, similar to that observed in DMD patients, was seen for CK and MYOM3. Sample size calculations suggested that low cohort sizes (N≤3) could be used to detect up to a 50% improvement in DE50-MD results towards WT levels for each biomarker or a combination thereof (via principal component analysis); as few as N=3 animals should enable detection of a 25% improvement using a combined biomarker approach (alpha 0.05, power 0.8). We have established a panel of blood-borne biomarkers that could be used to monitor musculoskeletal disease or response to a therapeutic intervention in the DE50-MD dog using low numbers of animals. The blood biomarker profile closely mimics that of DMD patients, supporting the hypothesis that this DMD model would be suitable for use in pre-clinical trials.
杜兴氏肌肉营养不良症（DMD）是一种由肌营养不良蛋白基因突变引起的致命性肌肉萎缩疾病。由于其与人类患者在表型上相似，大型动物模型是临床前试验中非常宝贵的工具。DE50-MD犬是一种相对较新的DMD模型，其携带的治疗性可处理突变位于人类患者的热点区域内，因此具有特别的价值。在使用这种新型动物模型进行治疗试验之前，建立一组可行的生物标志物至关重要。我们评估了DE50-MD犬中一组肌肉骨骼疾病的血源性生物标志物。在一项为期18个月的研究期间，每月从DE50-MD犬（N = 18）和野生型（WT）对照犬（N = 14）采集静脉血样。测量了一组DMD潜在的血浆/血清生物标志物，并使用样本量计算确定了它们在未来临床试验中的理论效用。与WT犬相比，DE50-MD犬的循环肌酸激酶（CK）活性、肌间蛋白-3（MYOM3）以及抗肌萎缩微RNA miR-1、miR-133a和miR-206显著更高，但血清肌生成抑制素浓度显著更低。CK和MYOM3呈现出与DMD患者中观察到的相似的年龄相关模式。样本量计算表明，对于每个生物标志物或其组合（通过主成分分析），低样本量（N≤3）可用于检测DE50-MD结果相对于WT水平高达50%的改善；使用联合生物标志物方法，仅N = 3只动物就应能够检测到25%的改善（α = 0.05，检验效能 = 0.8）。我们已经建立了一组血源性生物标志物，可用于使用少量动物监测DE50-MD犬的肌肉骨骼疾病或对治疗干预的反应。血液生物标志物谱与DMD患者的非常相似，支持了这一DMD模型适用于临床前试验的假设。
